Bruno Bost wrote:
>I'm looking for informations about studies on in vitro reconstructed
>metabolic pathways, used as tools to perform "in vitro" simulations
>on metabolic control, instead of "in silico" simulations.
>Does anyone hear about such studies ?
Johann's answer was a good one.
A very interesting set of examples on glycolysis is given by Robert K
Scopes' work in the 1970s (Biochem. J. 134, 197-208, 1973; 138, 119-123,
1973, and 42, 79-86, 1973, for example). He has reviewed this in a chapter
in the book: "New Beer in an Old Bottle: Eduard Buchner and the growth of
biochemical knowledge", ed. A Cornish-Bowden, University of Valencia, 1997.
A partial example is given by Nestor Tores and Enrique Melendez-Hevia, in
that they used a liver homogenate, but titrated it with purified enzymes
(hexokinase, phosphofructokinase) to determine the control coefficients:
Biochem. J. 234, 169-174, 1986. Martin Brand and Patti Quant used a
version of this experiment as a student practical at the University of
Cambridge.
As is clear in the case of Christoph Giersch's work, the problem of making
quantitative interpretations about conrol in the steady state is the
difficulty of achieving and maintaining a steady state in an in vitro system.
I have done some collaborative work with Jean Pierre Mazat in Bordeaux and
his students (Eric Quentin, Christophe Chassagnole) on an in vitro system
of E coli threonine synthesis, and it is easier to do dynamic studies on
such systems, which are also more informative for our purpose, which was to
build and test a model of the pathway, which can then be solved for steady
state conditions. Eric Quentin made heroic efforts to keep the system in a
steady state, but it's quite tricky, and essentially not posible if
something's going to change so you can also measure the flux.
David
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David Fell, School of Biological & Molecular Sciences,
Oxford Brookes University, Oxford OX3 0BP, UK
Tel. +44 (0)1865 483247 FAX +44 (0)1865 484017
http://www.brookes.ac.uk/bms/research/molcell/fell/mca_rg/mca_home.html
Understanding the control of metabolism: Spanish edition (Bases del
Control del Metabolismo) just published by Ediciones Omega, Barcelona.
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